Assay Method Information

Assay Name:  Alpha2-Adrenergic Receptor Binding Assay
Description:  The membrane preparation was prepared as described by Newman-Tancredi, A.; Nicolas, J,-P.; Audinot, V.; Gavaudan, S.; Verriele, L.; Touzard, M.; Chaput, C.; Richard, N.; Millan, N. J. (Action of alpha2 Adrenoreceptor Ligands at alpha2A and 5-HT1A Receptors: the Antagonist, Atipamezole, and the Agonist, Dexmedetomidine, are Highly Selective for alpha2A Adrenoreceptors. Naunyn-Schmiedeberg's Arch. Pharmacol. 1998, 358, 197-206). These membranes (30 ug proteins/ml for CHO-halpha2A and CHO-halpha2B, 100 ug protein/ml for CHO-halpha2C) were incubated for 60 min at ambient temperature in binding buffer (33 mM Tris-HCl, 1 mM EDTA, pH 7.5) in a final volume of 500 uL containing 0.8 or 1 or 2 nM [3H]RX821002 respectively for the adrenergic receptors halpha2A, halpha2B and halpha2C. Incubation was halted by rapid vacuum filtration through GF/C glass fibre filters followed by three successive washings in ice-cold binding buffer. Non-specific binding was determined using 10 uM phentolamine.
Affinity data for this assay
 

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